氯氰菊酯对小鼠小脑组织氧化损伤及核转录因子-E2相关因子2基因表达的影响

周礼华; 陈冲; 张朋; 王力; 吴学森

doi:10.13898/j.cnki.issn.1000-2200.2016.07.002

氯氰菊酯对小鼠小脑组织氧化损伤及核转录因子-E2相关因子2基因表达的影响

Effect of cypermethrin on the oxidative damage and mRNA expression of nuclear factor erythroid-2-related factor 2 in mice cerebellar tissue

摘要

摘要: 目的：研究氯氰菊酯对小鼠小脑组织氧化损伤及核转录因子-E2相关因子2（Nrf2）基因表达的影响。方法：以40只昆明小鼠为受试动物，随机分为4组，包括1个阴性对照组和3个氯氰菊酯染毒组，染毒组按5、10、20 mg/kg 3个剂量水平灌胃染毒小鼠3周，以小脑组织匀浆测定丙二醛（MDA）、谷胱甘肽过氧化物酶（GSH-PX）、总超氧化物歧化酶（T-SOD）、过氧化氢酶（CAT）的含量；用免疫组织化学法检测Nrf2在小脑组织细胞中的表达；用逆转录聚合酶链反应（RT-PCR）检测Nrf2基因表达。结果：随着氯氰菊酯染毒剂量升高，小脑组织MDA含量逐渐增加，GSH-PX、T-SOD、CAT等酶学活性逐渐下降，染毒组与对照组比较差异均有统计学意义（P<0.01）。免疫组织化学法显示，随着染毒剂量增加，小鼠小脑组织细胞核Nrf2阳性细胞表达增加，与对照组比较差异均有统计学意义（P<0.01），高剂量染毒组胞核阳性细胞表达均高于低剂量染毒组（P<0.01）。RT-PCR显示，随着染毒剂量增加，Nrf2条带光密度值增加，与对照组比较差异具有统计学意义（P<0.01），高剂量染毒组基因表达均高于低剂量染毒组（P<0.01）。结论：CYP染毒造成小鼠小脑组织氧化损伤，导致氧化应激反应中关键分子Nrf2由细胞质向细胞核转位，从而增强脑组织对抗氧化应激的能力。

Abstract: Objective: To observe the effects of cypermethrin on the oxidative damage and gene expression of nuclear factor erythroid-2-related factor 2(Nrf 2) mRNA in mice cerebellar tissue.Methods: Forty male Kunming mice were randomly divided into one negative control group and three cypermethrin infection groups.Three cypermethrin infection groups were infected by 5,10 and 20 mg/kg of cypermethrin gavage,once daily for three weeks,respectively.The contents of MDA,GSH-Px,T-SOD and CAT and MDA were measured in cerebellar tissue.The mRNA and protein expressions of Nrf2 in cerebellar tissue were detected using reverse transcription polymerase chain reaction(RT-PCR) and immunohistochemistry(IHC),respectively.Results: With the dose of cypermethrin increasing,the level of MDA increased,the levels of GSH-Px,T-SOD and CAT decreased in cerebellar tissue,and the differences of whose between the cypermethrin infection group and control group were statistically significant(P<0.01).The immunohistochemistry result showed that the positive expression of Nrf2 in nucleus increased in cerebellar tissue with the dose of cypermethrin increasing,the difference of which between the cypermethrin infection group and control group were statistically significant(P<0.01).The positive expression of Nrf2 in nucleus in high-dose cypermethrin infection group was higher than that in low-dose cypermethrin infection group(P<0.01).The RT-PCR results showed that the optical density value of Nrf2 increased in cerebellar tissue with the dose of cypermethrin increasing,the difference of which between the cypermethrin infection group and control group were statistically significant(P<0.01).The mRNA expression level of Nrf2 in high-dose cypermethrin infection group was higher than that in low-dose cypermethrin infection group(P<0.01).Conclusions: The cypermethrin infection can lead to the oxidative damage in cerebellar tissue and induce the translocation of Nrf2 from the cytoplasm to the nucleus,which can strength the antioxidative capacity of brain tissue.

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